1.
Pathological Investigations Of Theileriosis (T.Annulata) In Cattle In Disteict Lahore Punjab
by Syed Sadeed ud din Shah | Dr. Muti-ur- Rehman Khan | Dr. Raheela Akhtar | Prof. Dr. Aneela Zameer Durrani.
Material type: ; Format:
print
; Literary form:
not fiction
Publisher: 2014Dissertation note: Theileriosis is very important protozoal disease in crossbred cattle. According to an
assessment, about 250 million cattle are endangered by this disease and millions of high milk
yielding cattle are at risk of exposure to disease. It acts as a bigger restraint on livestock
improvement and production in many developing countries (Nagore et al. 2004). Theileria
annulata is the main specie that causes high morbidity and mortality. It causes heavy economic
and production losses in cattle in tropical and sub-tropical regions. The recorded mortality rates
in cattle reached to 70% (Moor house et al. 2001).
Theileria species are intracellular obligate hemoprotozoan parasites. All Theileria species
are dangerous and cause disease but two of them are important for livestock. Theileria parva and
T. annulata produces diseases named as East Coast fever and tropical theileriosis in cattle
respectively. Genus Theileria has many other species like T. buffeli, T. taurotragi, T. velifera, T.
sergenti and T. mutans. These species cause infections in wild and domesticated ruminants.
Theileria species present in large and small animals show signs like fever, anorexia,
swelling of the superficial lymph nodes, dyspnoea, lethargy, progressive anemia, constipation,
diarrhea, lacrimation and nervous symptoms (Saeed et al. 2010; Irvin and Mawmachi 1983). A
pronounced rise in body temperature, reaching 40-41.5 °C is pursued by lacrimation, depression,
swelling of the superficial lymph node and nasal discharge. The characteristic sign of tropical
theileriosis is anemia and finally haemoglobinuria occur with heavy weight losses. The clinical
course of the disease alter from per acute to acute or sub-acute to chronic (Oliveira- Sequeira et
al. 2005). The disease is lymphoproliferative in its early phases resulting enlargement of lymph
nodes, later on enters lymph destructive phase which is associated with a pronounced
Introduction
2
leukopenia. In the piroplasms phase in erythrocytes, the parasite becomes infective for the tick
(El-Deeb and Younis 2009).
Trans placental Bovine Tropical Theileriosis causing a deadly disease in a 3 day old
neonate cross bred calf and cerebral form of the disease (turning sickness) in a cow were
incriminated to T.annulata infection. It mainly depends upon the harmful effects of the T.
annulata on lymphoid tissues and susceptibility of the host (Sudan et al. 2012).
Theileriosis is prevalent in various regions of the world including Pakistan. It is transmitted by
Hyalomma species ticks. These ticks spread T. annulata which causes tropical theileriosis
(Durrani et al. 2009). The developmental stages of Theileria inside the Hyalomma ticks varies in
different shapes and forms (Hamed et al. 2011). Therefore to increase the milk and meat
production of cattle we can prevent the spread of the disease by controlling ticks
(Hekmatimoghaddam et al. 2012). The sufficient amount of Hyalomma ticks are found in warm,
commonly hard marshland and in central and Southern Europe, south west Asia and Southern
Africa having very long dry season. A toxin is produced in the adult ticks. This toxin produce
clinical signs of mucus membrane hyperemic and moist profuse eczema (Adam et al. 2000).
The sporozoites of Theileria enter into cattle host during tick feeding and they
immediately infect mononuclear leukocytes, these sporozoites develop into macroschizonts and
induce proliferation of the host cells. Macroschizonts constantly mature into microschizonts and
finally into merozoites, which are discharged from leukocytes. These merozoites attack
erythrocytes and mature into piroplasms, become available to ticks. Infective sporozoites,
injected during tick feeding, rapidly enter target cells, escape from the surrounding host-cell
membrane and differentiate to schizonts that interact with different host-cell components
(Dobbelaereand Rottenberg 2003). This interaction includes host cell signaling pathways that
Introduction
3
regulate proliferation and cell survival (Chaussepied and Langsley 2011) and thus cause
blastogenesis and clonal expansion of predominantly T and B cells (Fawcett et al. 1982; Baldwin
et al. 1988; Spooner et al. 1989). Merozoites released from these schizonts subsequently infect
red blood cells and become trophozoites. Lymphocytic stage of Theileria (schizonts) is the cause
of many of the severe disease manifestations like lymphadenopathy, pyrexia, thrombocytopenia,
and panleukopenia (Homer et al. 2000). Marked anemia, anisocytosis, pikilocytosis, and
leucopenia were commonly observed in bovine theileriosis (Ceci et al. 1997). Cattle may survive
the disease, but recovery and convalescence may be protracted and incomplete, this leads to
permanent debilitation, loss of productivity and prolonged carrier state. (Shahnawaz et al. 2011).
Cattle with subclinical infection in endemic regions become carrier of piroplasms and act as a
source of infection for the vectors (Brown 1997; Brown 1990; Uilenberg 1995).
The diagnosis of theileriosis in acute cases is majorly done on clinical signs and Giemsa
stained blood smears of cattle but the detection of agent is not reliable and is almost impossible
in carrier stage. Advances in molecular biological techniques have resulted in the improved
detection, identification, and genetic characterization of many hemoparasites. Species specific
polymerase chain reaction (PCR) has been developed for the detection and identification of
various Theileria species and has been shown to have higher sensitivity and specificity compared
with serological assays and examination of Giemsa-stained blood smears (Bhoora et al. 2009).
Primers were derived from the gene encoding the 30-kDa major merozoite surface antigen for T.
annulata (Aktas et al. 2006).
Most of the previous studies on haematological parameters in T. annulata infection were
carried out on experimentally infected cattle (Sandhu et al. 1998; Singh et al. 2001). The present
investigation was conducted to study haematological parameters in cattle naturally infected with
Introduction
4
T. annulata. Hematology has been broadly used in attempts to give information about disease
condition, performance problems and health in cattle (Rezaei and Naghadeh. 2006).
Hematological and sero-biochemical alterations are the indicators of severity of disease and are
considered to be good tools for the diagnosis, prognosis for effective therapy (Col and Uslu
2007; Nazifi et al. 2010b).
Lahore is one of the larger district in the Punjab province of Pakistan. Different cattle
breeds are reared by the people of the area for meat and milk production. The exact current
situation about the prevalence and pathogenesis of Theileriosis in the selected area is unknown.
The present study was conducted to screen cattle by finding schizonts or piroplasms in Giemsa
stained thin blood smears at slaughter house of district Lahore (Aktas et al. 2006) and later to
confirm through Polymerase Chain Reaction (PCR) (Chaisi et al. 2013) in order to implement
efforts and regulation to eradicate the spread of disease in the selected area. Data generated from
this study provided the latest status of Theileriosis, sex wise prevalence and its pathogenesis in
cattle population of Lahore. The study has also provided the necessary information to formulate
strategies for control of disease in the area. An investigation was also undertaken to ascertain the
changes in haematology as a result of Theileria annulata infection. These studies will help better
understanding of the pathogenesis and supportive therapy of this disease. Availability: Items available for loan: UVAS Library [Call number: 2186,T] (1).
2.
Common Nosocomial Bacterial Isolation And Identification From Veterinary Hospitals
by Muhammad Umar Zafar Khan (2008-VA-255) | Prof. Dr. Aneela Zameer Durrani | Prof.Dr.Muhammad Sarwar Khan | Dr. Hassan Bin Aslam.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: CD not available. Availability: Items available for loan: UVAS Library [Call number: 2217-T] (1).
3.
Comparison Of Two Imported Live Attenuated PPR Vaccines In Local Sheep In Pakistan
by Saliha Saba | Prof. Dr. Aneela Zameer Durrani | Dr. Hassan Salem | Dr. Imran Altaf.
Material type: ; Literary form:
not fiction
Publisher: 2015Dissertation note: Peste des petits ruminants (PPR) also famous as goat plaque is of viral origin and is extremely contagious disease of sheep and goat (Dhar et al. 2002; Asim et al. 2009). PPR can cause high mortality about 50 – 80 % in non-immunized sheep and goat population. Due to its similarity with other diseases, Peste des petits ruminants (PPR) is being devalued but at the same time it is said to be one of the major constraints to successful small ruminant farming in tropics (Sen et al. 2010). PPR virus is paramyxovirus, enveloped and belongs to the genus morbillivirus. These viruses comprise of 16Kb long, single stranded RNA showing negative polarity (Barrett et al. 2005).
The various vaccines like homologous and recombinant vaccines have been manufactured for the management of Peste des petits ruminants (PPR), as no accurate treatment is available for its control. For the immunity of animals against this disease, the tissue culture based, attenuated rinderpest vaccine (TCRV) had been accustomed over a extensive period because of the antigenic association among RPV and PPRV (Diallo et al. 1989).With the help of fresh freeze-drying methods and stabilizing agents the thermostability of the present PPR homologous vaccine has been enhanced significantly (Worrwall et al. 2001).
In Pakistan, PPR vaccine was manufactured with the help of PPRV Nigerian 75/I (PPR 75/1 LK 6 Vero 75) for the sheep and goat immunization (Asim et al. 2009). India had manufactured numerous live attenuated vaccines like the PPRV Sungri/96 that has been regularized for use (Hegde et al. 2008). ). The Peste des Petits Ruminants (PPRV-Sungri/96 ) vaccine is being manufactured on small and large scale for prevention of Peste des Petits Ruminants (PPR) outbreaks in India (Singh et al. 2004).
Summary
41
The current study was designed to study the immunogenicity of two imported live attenuated PPR vaccines in local sheep. A total of sixty (60) animals were selected and further separated into two groups, viz. Group-A and Group-B, having thirty (30) animals each. Group-A was further sub-divided into A1 comprising 10 sheep to which Raksha PPR vaccine (Sungri 96) was administered, A2 comprising of 10 sheep to which PPR vaccine (Nigeria 75/1) was administered and A3 comprising of 10 non-vaccinated sheep which served as control. Group B was separated into two sub-groups i.e B1 and B2 having fifteen (15) animals each. The Group-B1 was sub-divided into B1a having 05 sheep to which Raksha PPR vaccine (Sungri 96) was only administered, B1b having 05 sheep to which along with Raksha PPR vaccine (Sungri 96), Vitamin AD3E was administered and B1c having 05 unvaccinated sheep which served as control.
Similarly the Group-B2 was sub-divided into B2a having 05 sheep to which PPR vaccine (Nigeria 75/1) was only administered, B2b having 05 sheep to which along with PPR vaccine (Nigeria 75/1), Vitamin AD3E was administered and B2c having 05 non-vaccinated sheep and served as control group respectively. The serum samples were collected and mean antibody titer was calculated by complement fixation test (CFT) at zero day, 7th day, 14th day, 28th day and 48th day post-vaccination.
The live attenuated, Raksha PPR (Sungri 96) vaccine induced the mean antibody titers of 0 ±0.00, 4.7±0.48, 4.7±0.48, 4.9±0.31 and 4.9±0.31 which was significantly higher than the mean antibody titers shown by the PPR (Nigeria 75/1) vaccinated animals i.e. 0±0.00, 3.3±0.51, 3.4±0.51, 4±1.15 and 4.1±1.19 at zero, 7th, 14th, 28th, 48th day post-vaccination respectively. Similarly the mean antibody titers shown by the PPR (Nigeria 75/1) vaccinated animals were 0 ±0.00, 10.4± 3.86, 11.2±4.13, 20±11.31 and 21.6±11.80 at zero, 7th,14th, 28th and 48th day post vaccination respectively. Result of present study demonstrated
Summary
42
that the mean antibody titer values of animals vaccinated with Raksha PPR (Sungri 96) was significantly higher than animals vaccinated with PPR (Nigeria 75/1) at zero, 7th,14th, 28th and 48th day post vaccination respectively. The study also concluded that the mean antibody titer of animals receiving vaccination along with vitamin supplementation was significantly higher than animals receiving only vaccination. While performing the statistical analysis of data, it was revealed that the results were significant (p<0.05).
The present study summarized and concluded that the mean antibody titer values of Raksha PPR (Sungri 96) was significantly higher than PPR vaccine (Nigeria 75/1). As both India and Pakistan are two neighbouring countries, so PPR among them also falls in trans-boundary disease category. It signifies that both being part of Asia subcontinent and PPRV strain of lineage IV prevails in both regions. Keeping these factors under consideration proper vaccination strategy should be followed for the immunization of animals. In past, Nigeria 75/1 strain of PPRV vaccine had been used in Pakistan but the results were not reliable in terms of desired immune response and protection. Although titer was shown by this vaccine but protection is not reliable for proper health care of small ruminants. There was an immense need to come up with the authentic research on PPRV vaccine Raksha PPR (Sungri 96) in Pakistan which is already being used in India with desirable results.
The results of present research project were mostly similar with the findings of other scientists. The results of this study were analyzed through Independent t-test for independent samples. Availability: Items available for loan: UVAS Library [Call number: 2385-T] (1).
4.
Evaluation Of Risk Factors And Molecular Diagnosis Of Dermatophytosis In Dogs
by Muhammad Haseeb Saeed (2008-VA-241) | Prof. Dr. Aneela Zameer Durrani | Dr. Hassan Saleem | Prof. Dr. Azhar Maqbool.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Dogs are most kept and beloved pets in Pakistani society. Dermatophytosis is among the common disease of the pets. Many predisposing factors are involved in development of clinical cases of dermatophytosis including climatic conditions, housing condition of dogs and physical attributes such as coat hair size. Dermatophytosis is not only of concern as being infection of pets but also of its zoonotic importance hence it is very crucial to diagnose dermatophytic infection well in time. Dermatophytosis is caused by Dermatophytes,Microsporum, Trichophyton and Epidermophyton, the fungal species. It is difficult to diagnose the Dermatophytosis from other skin infections by routine tests in most of the cases especially subclinical. Polymerase Chain Reaction (PCR) is advanced and the most reliable technique to detect genome of Dermatophytes even in minute quantities specifically and can efficiently detect the presence of any Dermatophyte specie on the skin of dog. The current study was planned to develop and validate a diagnostic assay which could be able to detect and distinguish tree important dermatophytes species including Microsporum, Trichophyton and Epidermophytonby a uniplex PCR reaction. Analysis of involvement of certain predisposing factors in dermatophytosis was second goal to be worked on in this study. Samples of suspected pet dogs (n=50) were collected by scraping the skin at affected areas over skin. DNA was extracted from the skin scraping samples by organic Phenol Chloroform Isoamyle Alcohol method. Primers, specific to the 18-S ribosomal RNA region of genomes of the Dermatophytes, were designed after alignment of available sequences of Microsporum,Trichophyton and Epidermophyton at NCBI. Annealing temperature and recipe of PCR reaction was optimized by gradient PCR in BIO-Rad thermal cycler. Amplification reaction of all samples collected was carried out as per optimized reaction conditions, afterwards. Amplified products obtained were subjected to genotyping by agarose gel electrophoresis for size based separation of the amplified products. The specific amplified bands of desired genomic region of dermatophytes were seen in UV light transilluminator. The data of results of predisposing factors involved in dermatophytosis wasanalysedby using Pearson’s chi squared test with the help of Statistical Package for the Social Science (SPSS) Program.
Genome specific product sizes of Microsporum and Trichophyton i.e. 366 bp and 351 bp in respective positive samples were observed. Out of 50 suspected samples 46 samples were positive for dermatophytosis out of which 38 samples (82.6%) were positive for Microsporum, 6 samples (13%) for Trichophyton and 2 samples (4.4%) were positive for both Microsporumand Trichophyton.
This study will help to validate a diagnostic technique for Dermatophytosis with greater efficacy and reliability. Moreover, this investigation may become basis for the future research activities in this field in Pakistan.
Availability: Items available for loan: UVAS Library [Call number: 2528-T] (1).
5.
Study On Prevalence Of Intestinal Nematodes In Buffalo And Comparative Efficacy Of Herbal And Conventional Drugs Against Intestinal Nematodiosis In Buffalo Calves
by Abdul Rehman Qureshi (2014-VA-09) | Prof. Dr. Kamran Ashraf | Dr. Imran Rasheed | Prof. Dr. Aneela Zameer Durrani.
Material type: ; Literary form:
not fiction
Publisher: 2016Dissertation note: Parasites are a major cause of disease and production losses in livestock, frequently causing major economic loss and impacting on animal health. In livestock roundworms are deliberated the important economically upsetting internal parasites. Although buffalo can be diseased with tapeworms and flat worms, their effect on animal performance is minimal compared to the round worms. Buffalo feed on dry concentrate are less infected with parasite as compared to those who fed on pastures. The timing and frequency of anthelmintic treatments under such climatic conditions will vary greatly from place to place. Humid climates are permanently favorable for the development of infective larvae. With the frequently use of one or more dewormer animal produces anthelmintic resistance also its cost a lot for large herds. Herbal medicine is better option for control of intestinal worms. Herbal drugs are cheap, easy available and easy in administration.
The present study was designed to
1. Check the prevalence of intestinal nematodes in the buffalo population.
2. To determine the efficacy of herbal drugs against intestinal Nematodes in buffalo calves
3. To evaluate the comparison of effectiveness between herbal and commercial drugs to control the intestinal Nematodiosis in buffalo calves.
One hundred buffalo’s faecal samples were examined to check intestinal nematodes prevalence. All buffaloes belonging to various breeds were examined. 56 (93.33%) were Mix breed, 13 (72.22%) Nili Ravi breed and 18 (81.82%) of Kundi breed were positive for intestinal nematodiosis. 47
SUMMARY
Among the examined (87 %) buffaloes were found positive for nematodiosis. Ten buffaloes found positive (10 %) were less than 12 months, 25 buffaloes were between 1-2 year, 39% buffaloes were between 2 - 4 year and 13 buffaloes were above 4 years old were found positive for intestinal nematodiosis.
Among these buffaloes, 65 were male and 35 females. Out of male buffaloes 55 were found positive. While among 35 females 32 were positive for intestinal nematodes.
A total of 30 infected buffalo calves, of various ages (8-12 months), both sexes average body weight of 100 kg and naturally infected with intestinal nematodiosis were used for anthelmintic trials. These were randomly divided into 3 groups i.e. A, B, and C each having 10 calves. Group A and B were treated with herbal medicine. Group A was treated with dried powder Nigella Sativa (Kalonji) seeds at dose level of 250 mg/kg body weight. Group B was treated with Citrullus colocynthis (Kor Tumbha) fruit dried powder at dose level of 250 mg/kg body weight, group C was treated with Albandazole at dose level of 7.5 mg / kg body weight. The sample were taken on 0 day, 7th day, 14th, 21st and 28th day and EPG was determined by modified McMaster technique.
Statistical analysis was done using the statistical package for social science, (SPSS) version 20 (Chicago IL, USA). Data was presented as (mean+ S.D), the group descriptive measures were compared by CR Design (Anova) and applying differences were considered significant at P < 0.05. Post hoc test using Duncan multiple range test, to check the pair wise differences and alpha M. There was highly significant difference between commercial and herbal drugs. There was small significant difference between herbal drugs Nigella sativa and Citrullus 48
SUMMARY
colocynthis, both were highly effective against intestinal nematodiosis but less effective than Albandazole.
It is observed Nigella sativa dose showed a significant reduction in EPG .p value is p > .2284 at o day and at 7th day P>0.0146 ,at 14th day p> .0029 . There was also a significant decrease in EPG by Citrullus colocynthis but when compared these herbal drugs with commercial drugs the efficacy of herbal drug is 100 % at 21st day.
Recommendation.
Both herbal drugs Nigella sativa and Citrullus colocynthis used in trial were found highly effective against intestinal nematodiosis in buffalo calves but less effective than synthetic drug Albandazole hence, these herbal drugs cannot be recommended to be used as routine deworming of animals at farm level.
Keeping in view effectiveness of these herbal drugs in early age and having no side effects, it is recommended to be administered as feed additive to enhance immune-potentiation, effective anthelmintic and liver tonic.
There is dire need to carry out more research with increased dose rate of these herbal drugs and also use in combination with other herbal as well as synthetic drugs to evaluate its synergistic effect so farmer can be benefited of its maximum potential. Availability: Items available for loan: UVAS Library [Call number: 2637-T] (1).
6.
Epidemiology Of Major Bacterial And Parasitic Causes Of Foal Diarrhea
by Ikramul Haq (2010-VA-60) | Prof. Dr. Aneela Zameer Durrani | Prof. Dr. Muhammad Sarwar Khan | Dr. Muhammad Hassan Mushtaq.
Material type: ; Literary form:
not fiction
Publisher: 2017Dissertation note: Present study was carried out in District Lahore and District Sargodha, Punjab province of Pakistan, from January, 2016 to December, 2016. The study was conducted to study the prevalence of Diarrhea in foals and to identify the major viral, bacterial and parasitic causes of diarrhea in foals in these districts. The foals that passed lose feces a least 4 to 5 times a day were considered diarrheic. The results showed that the prevalence of diarrhea was 72.8% in the foals. District wise prevalence showed that the prevalence of diarrhea in foals were 73.7% in district Sargodha and were 72% in District Lahore. According to the results the prevalence of diarrhea in male foals was 74% and in female foal were 72%. The diarrhea was more prevalent in donkeys at is 76.6% as compaired to horses which was 74.5%.
The viral (rotavirus), bacterial (Salmonella, Clostridium perfirengens and E. coli) and parasitic causes of diarrhea were identified by appropriate technique. The viral causes were diagnosed using ELISA technique. The bacteria were isolated by culturing and were confirmed by polymerase Chain Reaction (PCR). The parasitic causes studied using microscopic examination. To identify the cause of diarrhea 400 samples (200 from each district) were collected and processed for viral, bacterial and parasitic detection.
The results showed that 91.1% of the samples were positive for one or more infectious agents. District wise results showed that the prevalence of more or more than infectious agents were higher in district Lahore (95.5%) as compared to district Sargodha which was 87.5%. The isolation of one or more than one infectious agents were higher in males it is 92.7% while were low in females which was 90.5%. The results showed that the prevalence of one or more than one infectious agents were higher in horses (92.4%) in comparision with donkey which was 87.8%.
Experiment No. I: Investigation of Parasitic causes of Foal Diarrhea
Fecal samples were preserved in 10% formalin and transported to the laboratory for diagnosis of parasites. The fecal samples from foals suffering from diarrhea were processed by using following parasitological examination.
4. Direct microscopic examination
The sample negative with direct microscopic examination was examined using simple floatation examination.
5. Simple floatation examination
The sample negative with Simple floatation examination was examined by using sedimentation floatation technique.
6. Sedimentation floatation Technique
The sample negative by using Sedimentation technique was recorded as negative for parasites.
The results show that 340 (85%) out of 400 samples were positive for one or more than one endo-parasites. The prevalence of endo-parasites was higher in district Sargodha it is 87.5% as compared to district Lahore, which was 82.5% (Table No.7). Gastrodiscus Spp were the higher prevalent endo-parasite and 308 (77%) (Table No. 10) of the samples were positive for Gastrodiscus Spp while the lowest prevalent endo-parasite was Anoplocephala spp with (3) 0.75% prevalence (Table No. 12). other helmenth such as Dictyocaulus Spp. (22.5%), Oxyuris Spp. (15.75%), Strongyloides Spp. (15.75%), Ascaris equorum (4.75), Tridontophorus Spp. (2%), Trichomena spp. (1.5%) Strongylus spp. (1.5%), and Paranoplocephala Spp. (5%)
Experiment No. II: Molecular Diagnosis of Bacteria Causes of Foal Diarrhea
The samples were culture for Salmonella, E.coli and Clostridium perfirengins on respective selective media and DNA was extracted from the culture. DNA was amplified by PCR and the bacteria were confirm using PCR. To diagnose Lasonia the DNA was extracted directly from fecal sample and were processed for lawsonia. The result show that 55% of the samples were positive for one or more than one type of bacteria. Maximum prevalence were observe of E. coli 48.75% and none of the sample were positive for lawsonia. The other isolated bacteria were Salmonella 18.24% and Clostridium perfiengens 18%.
Experiment No. III: Investigation of Viral causes of Foal Diarrhea
Foal suffering from diarrhea were screened and analyzed for presence of rotavirus by using commercially available ELISA kit
The result of detection of rotavirus shows that rotavirus was detected in (70) 17.5% of the sample processed for the diagnosis of rotavirus.
Availability: Items available for loan: UVAS Library [Call number: 2800-T] (1).
